The goal of the DNA Sequencing Shared Facility (DSSF) is to provide UAB Comprehensive Cancer Center (CCC) members with state-of-the-art automated Sanger sequencing. To this end, the specific aims are: 1. To continue to provide state-of-the-art automated DNA sequencing capabilities by maintaining dedicated Applied Biosystems Sequencers (ABI 3730XL) with capillary electrophoresis systems. 2. To continue to provide technical support and training to Cancer Center members, especially junior investigators, in DNA template preparation, primer design, cycle sequencing methodologies, and standard DNA sequence editing and analysis. To expand facility functions by providing microsatellite analyses and real-time PCR technologies as new services. To effectively interface with the Microarray Shared Facility (MSF) and the Comprehensive Genomics Shared Facility (CGSF) to assure services are complementary and not overlapping. Although originally established within the Center for AIDS Research (in 1987), CCC members now account for 70% of this facility's usage. This is because numerous laboratories within the Cancer Center are absolutely dependent on high throughput DNA sequencing for their research programs. The DSSF has ensured that CCC members have access to this service in a timely and cost-effective manner. In the last budget period, the DSSF has developed new methodologies for the automated sequencing process, trained CCC investigators in template preparation and the use of sequence analysis tools, upgraded and modernized pertinent equipment, employed automation technologies to expand capacity, and installed a Laboratory Information Management System to optimize the user interface. Since 2004, the DSSF has supported 104 CCC investigators and provided essential services for more than 175 cancer and cancer related grants and contracts. More than 458 million base pairs of primary sequence were determined, generating more than $2,710,000 in user charge-backs. Newly implemented automation increased the facility's efficiency and allowed the DSSF to reduce user charge-backs from $8 to $6 per sequencing reaction ($4 per reaction for 96 well plates).